Environmental detection of eumycetoma pathogens using multiplex real-time PCR for soil DNA in Sennar State, Sudan.

IF 3.6 Q1 TROPICAL MEDICINE Tropical Medicine and Health Pub Date : 2023-12-19 DOI:10.1186/s41182-023-00563-3
Hiroki Hashizume, Suguru Taga, Masayuki K Sakata, Mahmoud Hussein, Emmanuel Edwar Siddig, Toshifumi Minamoto, Ahmed Hassan Fahal, Satoshi Kaneko
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Abstract

Background: Mycetoma is a chronic disease affecting the skin and subcutaneous tissue endemic in the tropical and subtropical regions. Several bacteria and fungi can cause mycetoma, but fungal mycetoma (eumycetoma) is challenging because the treatment requires a combination of a long-term antifungal agent and surgery. Although the transmission route has not yet been elucidated, infection from the soil is a leading hypothesis. However, there are few soil investigation studies, and the geographical distribution of mycetoma pathogens is not well documented. Here, we used multiplex real-time PCR technology to identify three fungal species from soil samples.

Methods: In total, 64 DNA samples were extracted from soil collected in seven villages in an endemic area in Sennar State, Sudan, in 2019. Primers and fluorescent probes specifically targeting the ribosomal DNA of Madurella mycetomatis, Falciformispora senegalensis, and F. tompkinsii were designed.

Results: Multiplex real-time PCR was performed and identified the major pathogen, M. mycetomatis that existed in most sites (95%). In addition, two other pathogens were identified from some sites. This is the first report on the use of this technique for identifying the eumycetoma causative microorganisms.

Conclusions: This study demonstrated that soil DNA investigation can elucidate the risk area of mycetoma-causative agents. The results will contribute to the design of prevention measures, and further large-scale studies may be effective in understanding the natural habitats of mycetoma pathogens.

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利用苏丹森纳尔州土壤 DNA 的多重实时 PCR 技术,对埃米克特瘤病原体进行环境检测。
背景:霉菌瘤是一种影响皮肤和皮下组织的慢性疾病,是热带和亚热带地区的地方病。几种细菌和真菌都可引起霉菌瘤,但真菌性霉菌瘤(真菌瘤)的治疗具有挑战性,因为治疗需要结合长期抗真菌药物和手术。虽然传播途径尚未明确,但从土壤中感染是一个主要假设。然而,土壤调查研究很少,霉菌瘤病原体的地理分布也没有很好的记录。在此,我们利用多重实时 PCR 技术从土壤样本中鉴定了三种真菌:方法:2019 年,我们从苏丹森纳尔州一个地方病流行区的七个村庄采集的土壤中总共提取了 64 份 DNA 样本。设计了专门针对Madurella mycetomatis、Falciformispora senegalensis和F. tompkinsii核糖体DNA的引物和荧光探针:结果:进行了多重实时 PCR 检测,确定了主要病原体霉浆菌存在于大多数地点(95%)。此外,还在一些地点发现了另外两种病原体。这是第一份使用该技术鉴定真菌瘤致病微生物的报告:结论:这项研究表明,土壤 DNA 调查可以阐明霉菌瘤致病因子的风险区域。研究结果将有助于制定预防措施,进一步的大规模研究可能会有效了解霉菌瘤病原体的自然栖息地。
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来源期刊
Tropical Medicine and Health
Tropical Medicine and Health TROPICAL MEDICINE-
CiteScore
7.00
自引率
2.20%
发文量
90
审稿时长
11 weeks
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